Specificity and dynamics of transcriptional repression in retinal development

视网膜发育中转录抑制的特异性和动态

• 批准号: 9913544
• 负责人: Ilaria Rebay
• 金额: $ 20.25万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9913544
• 关键词: Address; Affinity; Alleles; Amino Acid Sequence; Binding Sites; Biochemical; Biological; Biological Assay; Biological Models; Biological Process; CRISPR/Cas technology; Cell physiology; Cells; Complex; DNA; DNA Binding; DNA Binding Domain; DNA Sequence; DNA-Protein Interaction; Defect; Development; Developmental Gene; Dimerization; Disease; Down-Regulation; Drosophila eye; Drosophila genus; ETV6 gene; Embryo; Engineering; Enhancers; Ensure; Family; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genomics; Goals; Heterogeneity; Human; In Vitro; MAP Kinase Gene; Mediating; Modeling; Mutation; Oncogenic; Orthologous Gene; Output; Phenotype; Phosphorylation; Photoreceptors; Positioning Attribute; Property; Protein Binding Domain; Proteins; Receptor Protein-Tyrosine Kinases; Regulation; Repression; Resolution; Retina; SAM Domain; Signal Transduction; Specific qualifier value; Specificity; Structure; System; Testing; Transcription Repressor; Visual system structure; Work; base; cell fate specification; computer studies; dimer; experimental study; fly; gain of function; gene repression; high risk; human disease; improved; in vivo; insight; leukemia; loss of function; multidisciplinary; mutant; nanomolar; progenitor; programs; protein protein interaction; response; transcription factor

! PROJECT SUMMARY Developmental programs are driven by the transcription factors that convert signaling information into the precise patterns of gene expression needed for accurate cell fate transitions. Transcription factor-DNA and transcription factor-transcription factor interactions together produce the necessary regulatory dynamics and precision. Because of the greater complexity and heterogeneity of protein-protein interactions, the field has focused on manipulating DNA sequence to study the mechanisms that determine enhancer specificity and gene expression output. This proposal describes our plan to manipulate transcription factor protein-protein interaction affinity. The experimental plan takes advantage of the deep mechanistic understanding and well- validated in vitro and in vivo assays established over two decades of studying the function and regulation of the conserved ETS family repressor Yan in the developing Drosophila visual system. Aim 1 outlines a multi- disciplinary strategy to engineer a set of yan alleles in which self-association affinity is gradually increased from weak to strong. Using these alleles, Aim 2 tests competing hypotheses for how protein-protein interaction affinity influences Yan function and target gene specificity during photoreceptor specification in the fly retina. Because the ideas we are testing address fundamental mechanisms of transcriptional and developmental regulation, the discoveries that emerge are likely to have impact well beyond our specific model system.

! 项目摘要 发育程序是由转录因子驱动的，转录因子将信号信息转化为细胞内信号。 精确的基因表达模式需要准确的细胞命运转换。转录因子-DNA和 转录因子-转录因子相互作用一起产生必要的调节动力学， 精度由于蛋白质-蛋白质相互作用的更大的复杂性和异质性，该领域已经 专注于操纵DNA序列来研究决定增强子特异性的机制， 基因表达输出。这个提议描述了我们操纵转录因子蛋白-蛋白的计划 相互作用亲和性该实验方案充分利用了深入的机理理解和良好的- 经过二十多年的研究， 在发育中的果蝇视觉系统中保守的ETS家族阻遏物Yan。目标1概述了一个多方面的 学科策略来工程化一组Yan等位基因，其中自缔合亲和力从 从弱到强使用这些等位基因，Aim 2测试了蛋白质-蛋白质相互作用如何相互竞争的假设。 亲合力影响果蝇视网膜感光细胞特化过程中Yan功能和靶基因特异性。 因为我们正在测试的想法解决了转录和发育的基本机制， 监管，出现的发现可能会产生远远超出我们特定模型系统的影响。